抗竹叶青蛇毒抗体F(ab′)2的制备

Preparation of F(ab′)2 fragment of Trimeresurus stejnegeri antibody

目的:制备特异性强、高纯度的抗竹叶青蛇毒抗体F(ab′)2。方法:以竹叶青蛇毒为免疫原免疫马,试血效价达到要求后,从免疫马血浆中先提取IgG,然后酶解、过疏水柱后得到F(ab′)2片段,以免疫扩散实验检测抗体F(ab′)2片段的特异性,以小鼠中和试验测定其中和竹叶青蛇毒的能力。结果:纯化得到的F(ab′)2活性片段纯度可达90%;F(ab′)2冻干品与竹叶青蛇毒在8∶1时出现免疫沉淀线,在20∶1时可保护小鼠100%存活。结论:纯化得到的抗体F(ab′)2活性片段对竹叶青蛇毒具有较好的中和能力。

Aim： To prepare the F（ab′）2 fragment of Trimeresurus stejnegeri antibody of high specificity and purity. Methods： IgG was extracted from the horse plasma immunized by the venom of Trimeresurus stejnegeri, then IgG was subjected to pepsin-digest, ammonium sulfate precipitation and chromatograph on hydrophobic column to obtain the F（ab′）2 active fragment. The specificity of antivenin was traced by way of immunodiffusion, and the protective effect was performed on mice in vitro. Results：The results of electrophoresis strip revealed that the purity of F（ab′）2 active fragment was up to 90%. The immune precipitation lines clearly appeared when the proportion of lyophilized antivenin and venom was 8：1; while it was 20：1, the mice are 100% survived in the neutralization test. Conclusion：The obtained antivenin can neutralize the venom efficiently.